用基因枪将GUS基因导入褐藻细胞中表达  

TRANSIENT EXPRESSION OF GUS GENE IN PHAEOPHYTES USING BIOLISTIC PARTICLE DELIVERY SYSTEM

作　　者：秦松[1] 张健[2] 李文斌[2] 王希华[1] 童顺 孙勇如[2] 曾呈奎[1] 

机构地区：[1]中国科学院海洋研究所,青岛266071 [2]中国科学院遗传研究所,北京100101

出　　处：《海洋与湖沼》1994年第4期353-356,共4页Oceanologia Et Limnologia Sinica

基　　金：国家科委项目;山东省科委攻关合同项目

摘　　要：于1993年11月-1994年2月，用高压氦气式基因枪，将PB1221质粒[装有CaMV35s启动子、GUS（β-葡精苷酸酶）基因以及nos的3’调控区]导入海带和裙带菜组织切块中。48h后，在海带假根细胞和裙带菜中肋部叶片细胞中检测到GUS基因的表达。实验表明，微粒子轰击法是外源基因导入大型褐藻的一个有效途径。CaMV35s启动子能够驱动外源基因在海洋藻类中的表达，可作为藻类基因工程的启动元件。Much progress has been made in recent years in transformation of blue-green algae and unicellular green algae. Both nuclear and chloroplast transformation have been achieved in Chlamydomonas. Only a little progress has been made in gene tran sfer of eukaryotic macroalgae, the seaweeds. Lack of an indirect transformation sys tem, such as Agrobacterium system, made the establishment of direct methods very important. We have little knowledge about promoters in seaweeds. Also, CaMV35S and SV40 have been successfully used in transformation of unicellular green algae.Levy (1991) reported preliminary trials of biolistics with Gracilaria but the results were negative. This paper reports study on transient expression of GUS gene in Laminaria japonica and Undaria pinnatifida using high velocity microprojectiles and CaMV35S as a useful promoter for seaweed transformation, and suggests an applica ble transformation mechanism capable of solving the regeneration problems of brown algal protoplasts.The materials used were healthy young sporophytes of L. japonica and U. pinnatifida collected from Qingdao. The middle parts of the blade and the rhizoidsof L. japonica were cut into 1×1cm2 and 1×0.3cml pieces separately. The bladeand the rhizoids of U. pinnatifida were cut the same way except that the coastaewere cut as new material (1×0.3cm2, or 1cm long and 0.8cm diameter). A BiolisticPDS-1000/He Particle Delivery System (Bio-Rad Company, U.S.A.) was used todeliver DNA into intact algal cells. Gold microprojectiles were coated with plasmidpB1221(CaMV35S promoter-GUS gene-nos ter). Control tissues were bombarded withgold microprojectiles without DNA. After two days' culture in MS medium, tissueswere stained for histochemical assay. Negative results were obtained for GUS activity in all control tissues. No GUS background reaction was found inside brown algal cells (Plate 1:1, 1:2) and blue spots were found scattered inside the rhizoids ofL. japonica (Plate 1:3, a cut-open rhizoid) and beneath the surface of costa of U.pinnatijid

关 键 词：褐藻 基因导入 GUS基因 细胞 

分 类 号：Q949.28[生物学—植物学]